Publication date: 25th July 2016
Synthetic scaffolds offer tremendous opportunities to manipulate cell behaviour and ultimately control neo-tissue formation and function during in situ tissue engineering. Yet, proper control over neo tissue formation requires in-depth understanding of cellular responses to complex environments. This is especially the case for cardiovascular tissue engineering, where cells are subject to combinations of structural (e.g. scaffold topography), biological (e.g. cytokines) and mechanical (cyclic stresses and strains) cues. Here, we report on the combined effects of topography and cyclic uniaxial strain on actin (re)orientation of vascular derived cells. In order to expose the cells to both cues, we created a modular setup of arrays of elastomeric microposts of different lengths (1, 3 and 6 mm, to vary substrate compliance), and cross sections (circular with radius of 1 μm radius and elliptical with major axis of 1.5 μm and minor axis of 0.87 μm), bound to a stretchable membrane to apply cyclic strain via a FlexCell system (7%, 0.5 Hz, sinus wave). Analysis of actin cytoskeleton orientation (immunocytochemistry, followed by image analysis) demonstrated a competition in response to topographical cues (contact guidance) and cyclic uniaxial strain (strain avoidance) when both cues were applied along the same direction. Interestingly, we observed that this competition originates from the distinct response of two subsets of actin stress fibers located respectively on top (actin cap) and below (basal) the nucleus. While basal fibers predominantly follow the topographical cues, actin cap fibers respond to the cyclic strain by strain avoidance. We also show that, upon uniaxial cyclic strain, nuclear orientation follows actin cap fiber orientation, suggesting that actin cap fibers play a major role in cellular and nuclear reorientation even in the presence of cell-aligning elliptical topographical cues. No influence of post length was observed. Repetition of the experiments using lamin A/C deficient cells, which lack a functional actin cap, confirmed the relevance of the actin cap in strain avoidance behaviour: the majority of these cells obey contact guidance only.
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